Volume 87, Issue 4, Pages 709-719 (November 1996) Loss of Fibrinogen Rescues Mice from the Pleiotropic Effects of Plasminogen Deficiency  Thomas H Bugge, Keith W Kombrinck, Matthew J Flick, Cynthia C Daugherty, Mary Jo S Danton, Jay L Degen  Cell  Volume 87, Issue 4, Pages 709-719 (November 1996) DOI: 10.1016/S0092-8674(00)81390-2

Figure 1 Fib Deficiency Rescues Plg−/− Mice from the Wasting Syndrome (A) Typical appearance of a control mouse, a Plg−/− mouse, a Fib−/− mouse, and a Plg−/−/Fib−/− mouse at seven months of age. Note the severe wasting of the Plg−/− mouse, whereas the appearance of the Plg−/−/Fib−/− mouse is indistinguishable from the control and Fib−/− mice. (B) Plot of weight versus age of control mice (blue diamonds), Fib−/− mice (red squares), Plg−/− mice (green triangles), and Plg−/−/Fib−/− mice (gold circles). The weights of a cohort initially consisting of 44 control mice, 25 Plg−/− mice, 16 Fib−/− mice, and 23 Plg−/−/Fib−/− mice were measured at 1- to 2-week intervals. Bars indicate the standard deviation. Due to the high mortality associated with Plg deficiency, the number of Plg−/− mice included in the weight measurements decreased over time as follows: days 20–100, 25 decreasing to 19 mice; days 101–200, 19 decreasing to 8 mice; days 201–300, 8 decreasing to 1 mouse. Cell 1996 87, 709-719DOI: (10.1016/S0092-8674(00)81390-2)

Figure 1 Fib Deficiency Rescues Plg−/− Mice from the Wasting Syndrome (A) Typical appearance of a control mouse, a Plg−/− mouse, a Fib−/− mouse, and a Plg−/−/Fib−/− mouse at seven months of age. Note the severe wasting of the Plg−/− mouse, whereas the appearance of the Plg−/−/Fib−/− mouse is indistinguishable from the control and Fib−/− mice. (B) Plot of weight versus age of control mice (blue diamonds), Fib−/− mice (red squares), Plg−/− mice (green triangles), and Plg−/−/Fib−/− mice (gold circles). The weights of a cohort initially consisting of 44 control mice, 25 Plg−/− mice, 16 Fib−/− mice, and 23 Plg−/−/Fib−/− mice were measured at 1- to 2-week intervals. Bars indicate the standard deviation. Due to the high mortality associated with Plg deficiency, the number of Plg−/− mice included in the weight measurements decreased over time as follows: days 20–100, 25 decreasing to 19 mice; days 101–200, 19 decreasing to 8 mice; days 201–300, 8 decreasing to 1 mouse. Cell 1996 87, 709-719DOI: (10.1016/S0092-8674(00)81390-2)

Figure 2 Fib Deficiency Rescues Plg−/− Mice from High Mortality Survival data are shown for a prospective cohort of 44 control mice, 25 Plg−/− mice, 16 Fib−/− mice, and 23 Plg−/−/Fib−/− mice that could be followed for at least 320 days. See Table 2 for details on morbidity and mortality of the mice. Cell 1996 87, 709-719DOI: (10.1016/S0092-8674(00)81390-2)

Figure 3 Alleviation of Organ Pathology in Plg−/− Mice by Fib Deficiency Representative histological sections of the liver (A and B), rectum (C and D), glandular stomach (E and F), and lung (G and H) of Plg−/− mice (A, C, E, and G) and Plg−/−/Fib−/− mice (B, D, F, and H). (A) Pathological changes in the liver of a 14-week-old Plg−/− mouse. Arrows indicate necrotic hepatocytes trapped in fibrin-rich patches. (B) Section of a histologically normal liver from a 16-week-old Plg−/−/Fib−/− mouse. Arrowheads delineate normal portal-tract structures. No necrotic hepatocytes are observed. (C) Longitudinal section of the rectum of a 16-week-old Plg−/− mouse with rectal prolapse, showing extensive ulceration of prolapsed mucosa with surface exudate and reactive hyperplasia (arrowheads). Triple arrows indicate muscular layers prolapsed past the squamocolumnar junction (asterisk). (D) Longitudinal section of the rectum of a 13-week-old Plg−/−/Fib−/− mouse. The two sets of double arrows indicate normal alignment of anal structures at the squamocolumnar junction (asterisks). (E) Ulceration of the glandular portion of the stomach (arrows) of a 24-week-old Plg−/− mouse. (F) Section of histologically normal glandular stomach and proximal duodenum of a 9-week-old Plg−/−/Fib−/− mouse. The arrowhead indicates the glandular stomach, the arrow indicates Brunners glands underlying the duodenum, and the asterisk indicates duodenal villi. (G) Area of organization (arrows) in the lung of a 17-week-old Plg−/− mouse. The asterisks indicate a bronchiole. (H) Corresponding section of a histologically normal lung of a 12-week-old Plg−/−/Fib−/− mouse. The asterisk indicates a terminal bronchiole. Magnification bars in A and B = 35 μm; C and D = 350 μm; E–H = 100 μm. Cell 1996 87, 709-719DOI: (10.1016/S0092-8674(00)81390-2)

Figure 4 Fib Deficiency Alleviates Impairment of Incisional Skin-Wound Healing in Plg−/− Mice (A) An example of progress in repair processes in a control mouse, a Plg−/− mouse, a Fib−/− mouse, and a Plg−/−/Fib−/− mouse at days 5, 12, and 16 after surgical incision. At day 5, all wounds are of similar size. The Fib−/− and Plg−/−/Fib−/− wounds appear slightly darker due to accumulation of dried blood at the wound margins. At day 12, a small residual defect is present in the wound of the control, Fib−/−, and Plg−/−/Fib−/− mice. In contrast, the wound from the Plg−/− mouse is still large; the scab is lost, and the wound appears red due to protruding granulation tissue. At day 16, the wounds of the control, Fib−/−, and Plg−/−/Fib−/− mice have healed, and new fur has grown to cover the wounds. The wound of the Plg−/− mouse is covered by a hard, scaly surface that lacks epidermal coverage. (B) Plot of the percentage of mice healed versus time after surgical incision: control mice (squares; n = 6), Plg−/− mice (triangles; n = 5), Fib−/− mice (circles; n = 6), and Plg−/−/Fib−/− mice (diamonds; n = 8). Cell 1996 87, 709-719DOI: (10.1016/S0092-8674(00)81390-2)

Figure 4 Fib Deficiency Alleviates Impairment of Incisional Skin-Wound Healing in Plg−/− Mice (A) An example of progress in repair processes in a control mouse, a Plg−/− mouse, a Fib−/− mouse, and a Plg−/−/Fib−/− mouse at days 5, 12, and 16 after surgical incision. At day 5, all wounds are of similar size. The Fib−/− and Plg−/−/Fib−/− wounds appear slightly darker due to accumulation of dried blood at the wound margins. At day 12, a small residual defect is present in the wound of the control, Fib−/−, and Plg−/−/Fib−/− mice. In contrast, the wound from the Plg−/− mouse is still large; the scab is lost, and the wound appears red due to protruding granulation tissue. At day 16, the wounds of the control, Fib−/−, and Plg−/−/Fib−/− mice have healed, and new fur has grown to cover the wounds. The wound of the Plg−/− mouse is covered by a hard, scaly surface that lacks epidermal coverage. (B) Plot of the percentage of mice healed versus time after surgical incision: control mice (squares; n = 6), Plg−/− mice (triangles; n = 5), Fib−/− mice (circles; n = 6), and Plg−/−/Fib−/− mice (diamonds; n = 8). Cell 1996 87, 709-719DOI: (10.1016/S0092-8674(00)81390-2)

Figure 5 Restoration of Keratinocyte Migration in Plg−/− Mice by Fib Deficiency Hematoxylin- and eosin-stained histological sections show a representative example of progress in wound repair in a control mouse (A and B), a Plg−/− mouse (C and D), a Fib−/− mouse (E and F), and a Plg−/−/Fib−/− mouse (G and H) 17 days after wounding. (A), (C), (E), and (G) are low magnifications showing the overall histological appearance of the wounds. (B), (D), (F), and (H) are high magnifications of the same sections demonstrating the progress in reepithelialization. By day 17, the keratinocyte wedges in the wounds from the control, Fib−/−, and Plg−/−/Fib−/− mice have met and fused to reepithelialize the wounds completely (arrows). Furthermore, in mice of each of these genotypes, granulation tissue has matured to form a scar. In contrast, at day 17, the wound from the Plg−/− mouse is still covered by a scab (asterisk in C and D), and the keratinocyte wedges (leading edges indicated by arrows) are near the wound margins. Magnification bars in (A), (C), (E), and (G) = 475 μm; (B), (D), (F), and (H) = 200 μm. Cell 1996 87, 709-719DOI: (10.1016/S0092-8674(00)81390-2)